Interstitial gamma-delta T cells in adriamycin-induced progressive glomerulosclerosis

T Ando1,2 H Wu1 DWatson1 T Hirano2 H Hirakata2 M Fujishima2 JF Knight1

1. Centre for Kidney Research, Royal Alexandra Hospital for Children, Westmead,
2. 2nd Dept of Internal Medicine. Kyushu University Fukuoka, Japan

While much is known about TCR gamma and delta gene organisation, our understanding of the pathophysiological role of cells bearing these receptors in renal disease is very limited. In this study 34 rats were given two injections of adriamycin (2.5 mg/kg BW) 14 days apart (ADR). Six controls were injected with saline. The adriamycin caused segmental glomerulosclerosis and massive interstitial infiltration of mononuclear cells with progression to end stage renal failure. Rats were sacrificed at 4 to 20 weeks after injection. Kidneys and lymph nodes were taken, the tissues were minced and mononuclear cells were isolated on a density gradient. Lymphocyte subpopulations were analysed with two-colour flow cytometry with antibodies to CD3, alpha-beta TCR and gamma-delta TCR. Renal tissues were fixed in 10% formalin and subjected to semiquantitative histology. Creatinine and urea were increased in ADR (Cr 4.8±2.8 vs 0.35±0.05, U 87±99 vs 19.1±3.4 mg/dl). Flow cytometry showed that gamma-delta T cells as a proportion of CD3+ cells were increased in ADR kidneys (8.5±2.8%) but not in abdominal lymph nodes (1.4±0.5%). The semiquantitative score of glomerular damage correlated significantly with the presence of gamma-delta T cells (r=0.65, p<0.01). Analysis of rat TCR V gamma repertoire by RT-PCR and Southern blotting showed that V gamma 2 was the predominant subfamily in peripheral lymph node tissue while V gamma 4 became the dominant subfamily in advanced stage rat ADR kidney. Sequencing of the N-region of V gamma 4 showed high junctional homogeneity. These studies suggest that gamma delta T cells participate in the renal tissue damage in this model and that a Vgamma 4+ gamma-delta T cell subpopulation may selectively recognise an unknown antigen.

Presented at the Australian and New Zealand Society of Nephrology, Brisbane, March 1999

Correspondence
Dr Huiling Wu
HuilW@chw.edu.au